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B6db references: nanana

type Journal Article
authors Kaulmann, U.; Smithies, K.; Smith, M.E.B.; Hailes, H.C.; Ward, J.M.
title Substrate spectrum of omega-transaminase from Chromobacterium violaceum DSM30191 and its potential for biocatalysis
journal Enzyme Microb Tech
Activity amine.transaminase
Family amine.transaminase
sel selected
ui nanana
year (2007)
volume 41
number 5
pages 628-637
 
keywords Biocatalysis; Chiral aminodiols; omega-Transaminase; alpha-Methylbenzylamine; Asymmetric synthesis
abstract A new omega-transaminase (ω-TA) from Chromobacterium violaceum DSM30191 has been recruited from the pool of fully sequenced genomes, using the V. fluvialis JS17 omega-TA sequence [Shin JS, Kim BG. Biosci Biotechnol Biochem 2001;65:17828; Shin JS, Yun H, Jang JW, Park I, Kim BG. Appl Microbiol Biotechnol 2003;61:46371] in a BLASTP search. The protein sequence of the C. violaceum enzyme exhibits 38% sequence identity to the V. fluvialis enzyme. The gene was functionally expressed in Escherichia coli BL21 by using the pET-expression system, making up 63% of the protein in the clarified cell extract (1.2 U/mg). As with the V. fluvialis TA, the C. violaceum enzyme shows (S)-enantioselective TA activity and converts (S)-greek small letter alpha-methylbenzylamine and pyruvate almost completely to acetophenone and l-Ala. Glyoxylate proved to be the best acceptor and aliphatic and aromatic aldehydes were both readily converted. Importantly we show here for the first time that an &omega-TA can be used for the amination of aromatic 2-keto-1,3-diols. An LC-ESI-MS method was developed to quantitatively detect aromatic aminodiol formation. Using clarified cell extract, we were able to convert 20 mM of racemic 1,3-dihydroxy-1-phenylpropan-2-one to 2-amino-1-phenylpropane-1,3-diol with 60% conversion in 24 h.
last changed 2011/04/13 13:09

B6db references